Immunoblotting Analysis of RIG-I Protein

Total proteins in cell lysate samples were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), followed by transfer to nitrocellulose membrane26 (link). Blotting of the membrane with antibodies against RIG-I (Santa Cruz Biotechnology, Inc., Dallas, TX) and tubulin (Sigma-Aldrich Corp, St. Louis, MO) was conducted. Horseradish peroxidase-conjugated secondary antibodies (Rockland Immunochemicals Inc.) and chemiluminescence substrate were used to reveal specific reactions by the primary antibodies. Chemi-Doc Imaging System (Bio-Rad, Hercules, CA) was used to capture the luminescence signal.

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Ma Z., Yu Y., Xiao Y., Opriessnig T., Wang R., Yang L., Nan Y., Samal S.K., Halbur P.G, & Zhang Y.J. (2016). Sustaining Interferon Induction by a High-Passage Atypical Porcine Reproductive and Respiratory Syndrome Virus Strain. Scientific Reports, 6, 36312.

Publication 2016

RIG-I Horseradish peroxidase-conjugated secondary antibodies Chemi-Doc Imaging System

Antibodies Cell Chemiluminescence Horseradish peroxidase Luminescence Membrane Nitrocellulose Proteins Rig i Sds page Tubulin

Corresponding Organization :

Other organizations : Virginia–Maryland College of Veterinary Medicine, University of Maryland, College Park, University of Edinburgh, Roslin Institute, Iowa State University

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Protein levels of RIG-I

control variables

Protein levels of tubulin

controls

Positive control: Blotting with antibodies against tubulin
Negative control: Not explicitly mentioned

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